WB result of Histone H3.3 Recombinant Rabbit mAb
Primary antibody: Histone H3.3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: Ramos whole cell lysate 20 µg
Lane 2: Raji whole cell lysate 20 µg
Lane 3: HeLa whole cell lysate 20 µg
Lane 4: PC-3 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Histone H3.3 Recombinant Rabbit mAb (S-3883)
Histone H3.3 Recombinant Rabbit mAb (S-3883)
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Product Details
Product Details
Product Specification
| Host | Rabbit |
| Antigen | Histone H3.3 |
| Synonyms | H3.3A; H3F3; H3F3A; H3-3A |
| Location | Nucleus |
| Accession | P84243 |
| Clone Number | S-3883 |
| Antibody Type | Recombinant mAb |
| Isotype | IgG |
| Application | WB, IHC-P, ICC, ChIP |
| Reactivity | Hu, Ms, Rt, Mk |
| Positive Sample | Ramos, Raji, HeLa, PC-3, NIH/3T3, mouse colon, rat stomach, COS-7 |
| Purification | Protein A |
| Concentration | 0.5 mg/ml |
| Conjugation | Unconjugated |
| Physical Appearance | Liquid |
| Storage Buffer | PBS, 40% Glycerol, 0.05% BSA, 0.03% Proclin 300 |
| Stability & Storage | 12 months from date of receipt / reconstitution, -20 °C as supplied |
Dilution
| application | dilution | species |
| WB | 1:1000 | Hu, Ms, Rt, Mk |
| IHC-P | 1:2000 | Hu, Ms, Rt |
| ICC | 1:500 | Hu |
| ChIP | 1:20-1:50 | Hu |
Background
Histone H3.3 is a replication-independent variant of the canonical histone H3 that can be deposited into nucleosomes throughout the cell cycle by dedicated chaperones such as HIRA, DAXX/ATRX, and DEK; it plays pivotal roles in gene activation, epigenetic memory, and chromatin remodeling because its distinct amino-acid sequence—especially residues that differ from H3.1/H3.2 like Ser31, Ala87, and Val89—alters nucleosome stability and creates preferential binding surfaces for transcription factors and histone-modifying enzymes, thereby influencing processes ranging from stem-cell pluripotency and development to neuronal activity and oncogenesis when its incorporation or mutations (e.g., the oncohistone Gly34Arg or Lys27Met) become dysregulated.
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Western Blot
WB result of Histone H3.3 Recombinant Rabbit mAb
Primary antibody: Histone H3.3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: NIH/3T3 whole cell lysate 20 µg
Lane 2: mouse colon lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
WB result of Histone H3.3 Recombinant Rabbit mAb
Primary antibody: Histone H3.3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: rat stomach lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
WB result of Histone H3.3 Recombinant Rabbit mAb
Primary antibody: Histone H3.3 Recombinant Rabbit mAb at 1/1000 dilution
Lane 1: COS-7 whole cell lysate 20 µg
Secondary antibody: Goat Anti-rabbit IgG, (H+L), HRP conjugated at 1/10000 dilution
Predicted MW: 15 kDa
Observed MW: 17 kDa
Immunohistochemistry
IHC shows positive staining in paraffin-embedded human colon. Anti-Histone H3.3 antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded human colon cancer. Anti-Histone H3.3 antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded mouse kidney. Anti-Histone H3.3 antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
IHC shows positive staining in paraffin-embedded rat stomach. Anti-Histone H3.3 antibody was used at 1/1000 dilution, followed by a HRP Polymer for Mouse & Rabbit IgG (ready to use). Counterstained with hematoxylin. Heat mediated antigen retrieval with Tris/EDTA buffer pH9.0 was performed before commencing with IHC staining protocol.
Immunocytochemistry
ICC shows positive staining in HeLa cells. Anti- Histone H3.3 antibody was used at 1/500 dilution (Green) and incubated overnight at 4°C. Goat polyclonal Antibody to Rabbit IgG - H&L (Alexa Fluor® 488) was used as secondary antibody at 1/1000 dilution. The cells were fixed with 4% PFA and permeabilized with 0.1% PBS-Triton X-100. Nuclei were counterstained with DAPI (Blue). Counterstain with tubulin (Red).
ChIP
Chromatin immunoprecipitation (ChIP) was performed on HeLa cells cross - linked with 1% formaldehyde for 10 min, then chromatin was fragmented by sonication. Parallel reactions used Histone H3.3 Recombinant Rabbit mAb (S-3883) and Rabbit mAb IgG Isotype Control (SDT-R173) at 1:50 for immunoprecipitation. Post - immunoprecipitation, both samples were washed, eluted, and cross - links reversed. Purified DNA was analyzed by qPCR.
qPCR showed the enrichment of RPL30, GAPDH, MYOD1,
AFM, SAT-α and SAT-2 in Histone H3.3 Recombinant
Rabbit mAb (S-3883)-immunoprecipitated
sample.
